Histone H4 (acetyl K16) Recombinant Antibody

$IF(ICC) staining with Histone H4 (acetyl K16) (2G2) Monoclonal Antibody (bsm-54330R) at 1:100 in Hela cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.$

IF(ICC) staining with Histone H4 (acetyl K16) (2G2) Monoclonal Antibody (bsm-54330R) at 1:100 in Hela cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.

$IF(ICC) staining with Histone H4 (acetyl K16) (2G2) Monoclonal Antibody (bsm-54330R) at 1:100 in SH-SY5Y cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.$

IF(ICC) staining with Histone H4 (acetyl K16) (2G2) Monoclonal Antibody (bsm-54330R) at 1:100 in SH-SY5Y cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.

Flow cytometric analysis of Hela cells with Histone H4 (acetyl K16) (2G2) Monoclonal Antibody (bsm-54330R) at a 1:100 dilution (red) compared with an unlabeled control (cells without incubation with primary antibody; black).

$The Hela (treated with Trichostatin A (TSA)(1\u03bcM, 18h)) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-Histone H4 (acetyl K16) antibody (bsm-54330R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.\r\n$

The Hela (treated with Trichostatin A (TSA)(1\u03bcM, 18h)) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-Histone H4 (acetyl K16) antibody (bsm-54330R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.\r\n

$4% Paraformaldehyde-fixed Hela (treated with Trichostatin A (TSA)(1\u03bcM, 18h)) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Histone H4 (acetyl K16)) monoclonal Antibody, unconjugated (bsm-54330R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.$

4% Paraformaldehyde-fixed Hela (treated with Trichostatin A (TSA)(1\u03bcM, 18h)) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Histone H4 (acetyl K16)) monoclonal Antibody, unconjugated (bsm-54330R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.

$Paraformaldehyde-fixed, paraffin embedded Mouse Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Esophageal Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Esophageal Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H4 (acetyl K16) Monoclonal Antibody, Unconjugated (bsm-54330R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Chromatin immunoprecipitation was performed using cross-linked chromatin from 5\u00d710\u2076 HeLa cells per IP, with 5 \u03bcg of bsm-54330R antibody or 5 \u03bcg of normal rabbit IgG(negative control bs-0295P), and 20 \u03bcL of Protein A\/G MagBeads per IP. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH-CDS and RPL30 exon 4 regions. The data are presented as enrichment of each sample relative to the total amount of input at each amplicon.$

Chromatin immunoprecipitation was performed using cross-linked chromatin from 5\u00d710\u2076 HeLa cells per IP, with 5 \u03bcg of bsm-54330R antibody or 5 \u03bcg of normal rabbit IgG(negative control bs-0295P), and 20 \u03bcL of Protein A\/G MagBeads per IP. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH-CDS and RPL30 exon 4 regions. The data are presented as enrichment of each sample relative to the total amount of input at each amplicon.

$C2C12 (M) treated with Trichostatin A (TSA) (1\u03bcM) for 18 h, HeLa (H) treated with Trichostatin A (TSA) (1\u03bcM) for 18 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with Histone H4 (acetyl K16) monoclonal antibody, unconjugated (bsm-54330R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

C2C12 (M) treated with Trichostatin A (TSA) (1\u03bcM) for 18 h, HeLa (H) treated with Trichostatin A (TSA) (1\u03bcM) for 18 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with Histone H4 (acetyl K16) monoclonal antibody, unconjugated (bsm-54330R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

Applications

WB
FCM
IHC-P
IF(IHC-P)
IF(ICC)

Reactivity

Human
Mouse
Rat

Overview
Catalog #	bsm-54330r
Product Name	Histone H4 (acetyl K16) Recombinant Antibody
Applications	WB FCM IHC-P IF(IHC-P) IF(ICC)
Reactivity	Human, Mouse, Rat
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Recombinant protein
Modification Site	K16
Clonality	Recombinant
Clone #	2G2
Isotype	IgG
Concentration	1mg/ml
Purification	purified by Protein A.
Storage Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	121504
Swiss Prot	P62805
Subcellular location	Nucleus
Synonyms	H4-16; H4/p; H4C1; H4C11; H4C12; H4C13; H4C14; H4C15; H4C2; H4C3; H4C4; H4C5; H4C6; H4C8; H4C9; HIST4H4; H4/o; H4C16; HIST2H4B; H4/m; H4FM; H4M; HIST1H4I; TEBIVANED4; TEVANED4; H4FA; HIST1H4A; H4/b; H4FB; HIST1H4D; dJ221C16.9; H4; H4/c; H4FC; HIST1H4F; H4/d; H4F2iii; H4FD; HIST1H4K; dJ160A22.1; H4/e; H4F2iv; H4FE; HIST1H4J; TEBIVANED2; TEVANED2; dJ160A22.2; H4/g; H4FG; HIST1H4C; TEBIVANED1; TEVANED1; dJ221C16.1; H4/h; H4FH; HIST1H4H; H4/I; H4FI; HIST1H4B; H4/j; H4FJ; HIST1H4E; TEBIVANED3; TEVANED3; H4.k; H4/k; H4FK; HIST1H4L; FO108; H4/n; H4F2; H4FN; HIST2H4; HIST2H4A; Gm11275; H4f16; Hist1h4m; Hist1h4n; B130044J01Rik; 1700024H08Rik; H2c8; H4_DROME; His4; His4r; His4:CG31611; His4:CG33869; His4:CG33871; His4:CG33873; His4:CG33875; His4:CG33877; His4:CG33879; His4:CG33881; His4:CG33883; His4:CG33885; His4:CG33887; His4:CG33889; His4:CG33891; His4:CG33893; His4:CG33895; His4:CG33897; His4:CG33899; His4:CG33901; His4:CG33903; His4:CG33905; His4:CG33907; His4:CG33909; H4r; H4_HUMAN; H4/A; H4FO; H4_MOUSE; H4-53; H4-12; H4_SCHPO; hhf1; hhf2; hhf3; h4.1; h4.2; h4.3; H4 clustered histone 1; H4 histone family, member A; histone 1, H4a; histone cluster 1, H4a; histone cluster 1 H4 family member a; H4C1 \| Histone H4 (acetyl-K16); H4K16Ac; ac-Histone H4; acetyl-Histone H4; ac-H4; H4C1 \| Histone H4 (acetyl-Lys16)
Background	Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Histone H4 are known to be acetylated on Lysine 5, 8, 12, and 16. Acetylations are important for the regulation of histone deposition, transcriptional activation, DNA replication and repair. Hyper-acetylation of the histone tails is shown to neutralize positive charge and weakens histone-DNA and nucleosome-nucleosome interactions that destabilizes chromatin structure and increases the accessibility of DNA to various DNA-binding proteins.
Application Dilution
WB	1:300-5000
FCM	1:20-100
IHC-P	1:200-400
IF(IHC-P)	1:50-200
IF(ICC)	1:50-200